不同宫颈癌临床分期患者外周血中T淋巴细胞及NK细胞的表达分析
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摘要
目的分析宫颈癌患者外周血中T淋巴细胞及其亚群以及NK细胞的表达情况。方法前瞻性分析145例在我院就诊的宫颈癌患者的临床资料,按照宫颈癌FIGO临床分期差别分为:宫颈癌Ⅰ期38例、宫颈癌Ⅱ期42例、宫颈癌Ⅲ期35例、宫颈癌Ⅳ期30例;同期选择体检正常女性30例作为对照组。应用流式细胞仪检测各组患者外周血CD3~+、CD4~+、CD8~+T细胞亚群、调节性T细胞(Treg)以及NK细胞数量,同时计算各亚群的比例以及Treg占CD4~+T细胞的比例。应用多元Logistic回归分析评估各细胞亚群数量及比例与宫颈癌临床分期的相关性。结果与对照组相比,宫颈癌患者CD3~+T细胞、CD4~+T细胞、NK细胞数量以及CD4~+/CD8~+比值均较低,Treg/CD4~+比值较高(均P <0.05),而CD8~+T细胞数量差异无统计学意义(均P>0. 05)。随宫颈癌病理严重程度递增,CD3~+T细胞、CD4~+T细胞、NK细胞数量以及CD4~+/CD8~+比值逐步降低,Treg/CD4~+比值逐步增高(均P <0.05),而CD8~+T细胞数量差异无统计学意义(均P>0.05)。多元Logistic回归分析显示,Treg/CD4~+比值是宫颈癌临床分期的危险因素,CD4~+T细胞、CD4~+/CD8~+比值及NK细胞是其保护性因素(均P<0.05)。结论宫颈癌患者细胞免疫功能均不同程度降低,晚期患者降低最显著。检测T淋巴细胞亚群及NK细胞可用于宫颈癌患者免疫监测,为临床治疗及预后评估提供参考。
Objective To analyze the expressions of T cell subsets and NK cells in peripheral blood cell of patients with cervical cancer. Methods The clinical data of 145 in-patients with cervical cancer were collected for the prospectively study. All patients were divided according to the stages of the International Federation of Gynecology and Obstetrics(FIGO), which were stage Ⅰ cervical cancer(38 cases),stage Ⅱ cervical cancer(42 cases),stage Ⅲ cervical cancer(35 cases) and stage IV cervical cancer(30 cases). And 30 cases of healthy women were enrolled as the healthy control. The levels of peripheral blood CD3~+, CD4~+, CD8~+ T lymphocyte subsets and regulatory T cells(Treg cells) and natural killer(NK) cells were detected by flow cytometry. And the ratios of CD4 ~+/CD8 ~+ and Treg/CD4 ~+ were calculated. The correlations of T lymphocyte subsets, NK cells and ratios with the stages of the FIGO were analyzed by multivariate logistic regression analysis. Results Compared with the healthy control, the levels of peripheral blood CD3~+, CD4~+ T lymphocyte subsets and NK cells and the ratios of CD4~+/CD8~+ in the patients with cervical cancers were lower, and the ratio of Treg/CD4~+ in the patients with cervical cancers was higher(all P <0. 05),but the level of peripheral blood CD8 ~+ T lymphocyte subsets were no different between the two groups(all P>0.05). With the increasing severity of cervical cancer lesions,the levels of peripheral blood CD3 ~+,CD4~+ T lymphocyte subsets and NK cells and the ratio of CD4~+/CD8~+ were gradually reduced, and the ratio of Treg/CD4~+ was gradually increased( all P < 0.05),but the levels of peripheral blood CD8~+ T lymphocyte subsets were no different between the two groups(all P > 0.05). The CD4~+ T lymphocyte subsets, NK cells and the ratio of CD4~+/CD8~+ were confirmed to be the protective factors for the stage of the FIGO and the ratio of Treg/CD4~+ was confirmed to be the risk factors by multivariate logistic regression analysis(P >0. 05).Conclusion The cellular immune functions of the patients with cervical cancer are lower and even particularly worse in patients with advanced clinical stages of cervical cancer. Detecting T lymphocyte subsets and NK cells can be used to monitor cellular immune function of patients with cervical cancer, so to estimate the prognosis and provide a reference for the clinical treatment.
Objective To analyze the expressions of T cell subsets and NK cells in peripheral blood cell of patients with cervical cancer. Methods The clinical data of 145 in-patients with cervical cancer were collected for the prospectively study. All patients were divided according to the stages of the International Federation of Gynecology and Obstetrics(FIGO), which were stage Ⅰ cervical cancer(38 cases),stage Ⅱ cervical cancer(42 cases),stage Ⅲ cervical cancer(35 cases) and stage IV cervical cancer(30 cases). And 30 cases of healthy women were enrolled as the healthy control. The levels of peripheral blood CD3~+, CD4~+, CD8~+ T lymphocyte subsets and regulatory T cells(Treg cells) and natural killer(NK) cells were detected by flow cytometry. And the ratios of CD4 ~+/CD8 ~+ and Treg/CD4 ~+ were calculated. The correlations of T lymphocyte subsets, NK cells and ratios with the stages of the FIGO were analyzed by multivariate logistic regression analysis. Results Compared with the healthy control, the levels of peripheral blood CD3~+, CD4~+ T lymphocyte subsets and NK cells and the ratios of CD4~+/CD8~+ in the patients with cervical cancers were lower, and the ratio of Treg/CD4~+ in the patients with cervical cancers was higher(all P <0. 05),but the level of peripheral blood CD8 ~+ T lymphocyte subsets were no different between the two groups(all P>0.05). With the increasing severity of cervical cancer lesions,the levels of peripheral blood CD3 ~+,CD4~+ T lymphocyte subsets and NK cells and the ratio of CD4~+/CD8~+ were gradually reduced, and the ratio of Treg/CD4~+ was gradually increased( all P < 0.05),but the levels of peripheral blood CD8~+ T lymphocyte subsets were no different between the two groups(all P > 0.05). The CD4~+ T lymphocyte subsets, NK cells and the ratio of CD4~+/CD8~+ were confirmed to be the protective factors for the stage of the FIGO and the ratio of Treg/CD4~+ was confirmed to be the risk factors by multivariate logistic regression analysis(P >0. 05).Conclusion The cellular immune functions of the patients with cervical cancer are lower and even particularly worse in patients with advanced clinical stages of cervical cancer. Detecting T lymphocyte subsets and NK cells can be used to monitor cellular immune function of patients with cervical cancer, so to estimate the prognosis and provide a reference for the clinical treatment.
引文
[1]李军,王一羽,原荣,等.高危型HPV感染者外周血T细胞亚群和NK细胞检测及临床价值[J].中华实验和临床病毒学杂志,2015,29(6):465-468.
[2] QI L,HE W. REGγis associated with lymph node metastasis and Tstage in papillary thyroid carcinoma[J]. Med Sci Monit,2018,24:1373-1378.
[3] LUCENA A A, GUIMARAES M V, MICHELIN M A, et al.Evaluation of T, B and natural killer lymphocyte in the cervical stroma of HIV-positive and negative patients with cervical intraepithelial neoplasia[J]. Immunol Lett,2016,169:98-103.
[4] XUE J,WANG Y,CHEN C,et al. Effects of Th17 cells and IL-17in the progression of cervical carcinogenesis with high-risk human papillomavirus infection[J]. Cancer Med,2018,7(2):297-306.
[5] ZHANG Y,ZHU W, ZHANG X, et al. Expression and clinical significance of programmed death-1 on lymphocytes and programmed death ligand-1 on monocytes in the peripheral blood of patients with cervical cancer[J]. Oncol Lett,2017,14(6):7225-7231.
[6] ZHANG T,JIAO J,JIAO X,et al. Aberrant frequency of TNFR2~+Treg and related cytokines in patients with CIN and cervical cancer[J]. Oncotarget, 2017,9(4):5073-5083.
[7] RAM B M, DOLPADY J, KULKARNI R, et al. Human papillomavirus(HPV)oncoprotein E6 facilitates Calcineurin-Nuclear factor for activated T cells 2(NFAT2)signaling to promote cellular proliferation in cervical cell carcinoma[J]. Exp Cell Res,2018,362(1):132-141.
[8] KOSKIMAA H M, PAASO A E, WELTERS M J, et al. Human papillomavirus 16 E2-, E6-and E7-specific T-cell responses in children and their mothers who developed incident cervical intraepithelial neoplasia during a 14-year follow-up of the Finnish Family HPV cohort[J].J Transl Med,2014,12:44.
[9] Da SILVA D M,WOODHAM A W,SKEATE J G,et al. Langerhans cells from women with cervical precancerous lesions become functionally responsive against human papillomavirus after activation with stabilized Poly-I:C[J]. Clin Immunol, 2015, 161(2):197-208.
[10] MALDONADO ALVARADO E,OSORIO PERALTA MO,MORENO VAZQUEZ A, et al. Effectiveness of photodynamic therapy inelimination of HPV-16 and HPV-18 associated with CIN I in Mexican women[J]. Photochem Photobiol, 2017, 93(5):1269-1275.
[11] PAPASAVVAS E,SURREY L F,GLENCROSS D K,et al. Highrisk oncogenic HPV genotype infection associates with increased immune activation and T cell exhaustion in ART-suppressed HIV-1-infected women[J]. Oncoimmunology,2016,5(5):e1128612.
[12] PAASO A,KOSKIMAA H M, WELTERS M J,et al. Cell mediated immunity against HPV16 E2,E6 and E7 peptides in women with incident CIN and in constantly HPV-negative women followed-up for 10-years[J]. J Transl Med,2015,13:163.
[13] McCORMACK S E,CRUZ C R Y, WRIGHT K E,et al. Human papilloma virus-specific T cells can be generated from na(?)ve T cells for use as an immunotherapeutic strategy for immunocompromised patients[J]. Cytotherapy,2018,20(3):385-393.
[14] BENARD V B, CASTLE P E, JENISON S A, et al. Populationbased incidence rates of cervical intraepithelial neoplasia in the human papillomavirus vaccine era[J]. JAMA Oncol,2017,3(6):833-837.
[15] KURMYSHKINA 0 V,KOVCHUR P I,SCHEGOLEVA L V,et al.T-and NK-cell populations with regulatory phenotype and markers of apoptosis in circulating lymphocytes of patients with CIN3 or microcarcinoma of the cervix:evidence for potential mechanisms of immune suppression[J]. Infect Agent Cancer,2017,12:56.
[16] SHRESTHA R L,AHN G S,STAPLES M I,et al. Mislocalization of centromeric histone H3 variant CENP-A contributes to chromosomal instability(CIN)in human cells[J]. Oncotarget, 2017,8(29):46781-46800.
[17] POROPATICH K,FONTANAROSA J, SWAMINATHAN S,et al.Comprehensive T-cell immunophenotyping and next-generation sequencing of human papillomavirus(HPV)-positive and HPVnegative head and neck squamous cell carcinomas[J]. J Pathol,2017,243(3):354-365.
[18] MICHELIN M A, MONTES L, NOMELINI R S, et al. Helper T lymphocyte response in the peripheral blood of patients with intraepithelial neoplasia submitted to immunotherapy with pegylated interferon-a[J].Int J Mol Sci,2015,16(3):5497-5509.
[19] UIJTERWAAL M H, van ZUMMEREN M, KOCKEN M, et al.Performance of CADM1/MAL-methylation analysis for monitoring of women treated for high-grade CIN[J]. Gynecol Oncol, 2016,143(1):135-142.
[20] VU LTH,TRAN H T D, NGUYEN B T,et al. Community-based screening for cervical cancer using visual inspection with acetic acid:results and lessons learned from a pilot study in Vietnam[J]. J Public Health Manag Pract,2018,24(Suppl 2):S3-S8.
[2] QI L,HE W. REGγis associated with lymph node metastasis and Tstage in papillary thyroid carcinoma[J]. Med Sci Monit,2018,24:1373-1378.
[3] LUCENA A A, GUIMARAES M V, MICHELIN M A, et al.Evaluation of T, B and natural killer lymphocyte in the cervical stroma of HIV-positive and negative patients with cervical intraepithelial neoplasia[J]. Immunol Lett,2016,169:98-103.
[4] XUE J,WANG Y,CHEN C,et al. Effects of Th17 cells and IL-17in the progression of cervical carcinogenesis with high-risk human papillomavirus infection[J]. Cancer Med,2018,7(2):297-306.
[5] ZHANG Y,ZHU W, ZHANG X, et al. Expression and clinical significance of programmed death-1 on lymphocytes and programmed death ligand-1 on monocytes in the peripheral blood of patients with cervical cancer[J]. Oncol Lett,2017,14(6):7225-7231.
[6] ZHANG T,JIAO J,JIAO X,et al. Aberrant frequency of TNFR2~+Treg and related cytokines in patients with CIN and cervical cancer[J]. Oncotarget, 2017,9(4):5073-5083.
[7] RAM B M, DOLPADY J, KULKARNI R, et al. Human papillomavirus(HPV)oncoprotein E6 facilitates Calcineurin-Nuclear factor for activated T cells 2(NFAT2)signaling to promote cellular proliferation in cervical cell carcinoma[J]. Exp Cell Res,2018,362(1):132-141.
[8] KOSKIMAA H M, PAASO A E, WELTERS M J, et al. Human papillomavirus 16 E2-, E6-and E7-specific T-cell responses in children and their mothers who developed incident cervical intraepithelial neoplasia during a 14-year follow-up of the Finnish Family HPV cohort[J].J Transl Med,2014,12:44.
[9] Da SILVA D M,WOODHAM A W,SKEATE J G,et al. Langerhans cells from women with cervical precancerous lesions become functionally responsive against human papillomavirus after activation with stabilized Poly-I:C[J]. Clin Immunol, 2015, 161(2):197-208.
[10] MALDONADO ALVARADO E,OSORIO PERALTA MO,MORENO VAZQUEZ A, et al. Effectiveness of photodynamic therapy inelimination of HPV-16 and HPV-18 associated with CIN I in Mexican women[J]. Photochem Photobiol, 2017, 93(5):1269-1275.
[11] PAPASAVVAS E,SURREY L F,GLENCROSS D K,et al. Highrisk oncogenic HPV genotype infection associates with increased immune activation and T cell exhaustion in ART-suppressed HIV-1-infected women[J]. Oncoimmunology,2016,5(5):e1128612.
[12] PAASO A,KOSKIMAA H M, WELTERS M J,et al. Cell mediated immunity against HPV16 E2,E6 and E7 peptides in women with incident CIN and in constantly HPV-negative women followed-up for 10-years[J]. J Transl Med,2015,13:163.
[13] McCORMACK S E,CRUZ C R Y, WRIGHT K E,et al. Human papilloma virus-specific T cells can be generated from na(?)ve T cells for use as an immunotherapeutic strategy for immunocompromised patients[J]. Cytotherapy,2018,20(3):385-393.
[14] BENARD V B, CASTLE P E, JENISON S A, et al. Populationbased incidence rates of cervical intraepithelial neoplasia in the human papillomavirus vaccine era[J]. JAMA Oncol,2017,3(6):833-837.
[15] KURMYSHKINA 0 V,KOVCHUR P I,SCHEGOLEVA L V,et al.T-and NK-cell populations with regulatory phenotype and markers of apoptosis in circulating lymphocytes of patients with CIN3 or microcarcinoma of the cervix:evidence for potential mechanisms of immune suppression[J]. Infect Agent Cancer,2017,12:56.
[16] SHRESTHA R L,AHN G S,STAPLES M I,et al. Mislocalization of centromeric histone H3 variant CENP-A contributes to chromosomal instability(CIN)in human cells[J]. Oncotarget, 2017,8(29):46781-46800.
[17] POROPATICH K,FONTANAROSA J, SWAMINATHAN S,et al.Comprehensive T-cell immunophenotyping and next-generation sequencing of human papillomavirus(HPV)-positive and HPVnegative head and neck squamous cell carcinomas[J]. J Pathol,2017,243(3):354-365.
[18] MICHELIN M A, MONTES L, NOMELINI R S, et al. Helper T lymphocyte response in the peripheral blood of patients with intraepithelial neoplasia submitted to immunotherapy with pegylated interferon-a[J].Int J Mol Sci,2015,16(3):5497-5509.
[19] UIJTERWAAL M H, van ZUMMEREN M, KOCKEN M, et al.Performance of CADM1/MAL-methylation analysis for monitoring of women treated for high-grade CIN[J]. Gynecol Oncol, 2016,143(1):135-142.
[20] VU LTH,TRAN H T D, NGUYEN B T,et al. Community-based screening for cervical cancer using visual inspection with acetic acid:results and lessons learned from a pilot study in Vietnam[J]. J Public Health Manag Pract,2018,24(Suppl 2):S3-S8.